DNA binding by the ribosomal DNA transcription factor rrn3 is essential for ribosomal DNA transcription.

The human homologue of yeast Rrn3 is an RNA polymerase I-associated transcription factor that is essential for ribosomal DNA (rDNA) transcription. The generally accepted model is that Rrn3 functions as a bridge between RNA polymerase I and the transcription factors bound to the committed template. In this model Rrn3 would mediate an interaction between the mammalian Rrn3-polymerase I complex and SL1, the rDNA transcription factor that binds to the core promoter element of the rDNA. In the course of studying the role of Rrn3 in recruitment, we found that Rrn3 was in fact a DNA-binding protein. Analysis of the sequence of Rrn3 identified a domain with sequence similarity to the DNA binding domain of heat shock transcription factor 2. Randomization, or deletion, of the amino acids in this region in Rrn3, amino acids 382-400, abrogated its ability to bind DNA, indicating that this domain was an important contributor to DNA binding by Rrn3. Control experiments demonstrated that these mutant Rrn3 constructs were capable of interacting with both rpa43 and SL1, two other activities demonstrated to be essential for Rrn3 function. However, neither of these Rrn3 mutants was capable of functioning in transcription in vitro. Moreover, although wild-type human Rrn3 complemented a yeast rrn3-ts mutant, the DNA-binding site mutant did not. These results demonstrate that DNA binding by Rrn3 is essential for transcription by RNA polymerase I.
Mesh Terms:
Amino Acid Sequence, Animals, Cell Line, Cell Nucleolus, DNA, DNA, Ribosomal, DNA-Binding Proteins, Gene Deletion, Humans, Insects, Mice, Molecular Sequence Data, Mutagenesis, Pol1 Transcription Initiation Complex Proteins, Promoter Regions, Genetic, Protein Binding, RNA, Ribosomes, Saccharomyces cerevisiae Proteins, Sequence Homology, Amino Acid, TATA-Binding Protein Associated Factors, Transcription Factor TFIID, Transcription, Genetic
J. Biol. Chem. Mar. 29, 2013; 288(13);9135-44 [PUBMED:23393135]
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